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Nitrate-limited batch cultures, incorporating 20 different fermentation substrates and inoculated with human faeces, mainly selected for the growth of enterobacteria. The microbial diversity involved was determined by a combination of phenotypic and genotypic procedures. Continuous culture with lactate as the sole electron donor selected for similar micro-organisms, but when antibiotics were incorporated to inhibit Escherichia coli and lactate was replaced with choline, there was a wider microbial diversity recovered. Clostridium ramosum and Bacteroides vulgatus were then isolated as well as enterobacteriaceae.  相似文献   

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Abstract: Different reduced sulfur compounds (H2S, FeS, S2O32−) were tested as electron donors for dissimilatory nitrate reduction in nitrate-amended sediment slurries. Only in the free sulfide-enriched slurries was nitrate appreciably reduced to ammonia (     ), with concomitant oxidation of sulfide to S0 (     ). The initial concentration of free sulfide appears as a factor determining the type of nitrate reduction. At extremely low concentrations of free S2− (metal sulfides) nitrate was reduced via denitrification whereas at higher S2− concentrations, dissimilatory nitrate reduction to ammonia (DNRA) and incomplete denitrification to gaseous nitrogen oxides took place. Sulfide inhibition of NO- and N2O- reductases is proposed as being responsible for the driving part of the electron flow from S2− to NH4+.  相似文献   

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The inhibitory effects of nitrate (NO3-) and nitrite (NO2-) on dissimilatory iron (FE3+) reduction were examined in a series of electron acceptor competition experiments using Shewanella putrefaciens 200 as a model iron-reducing microorganism. S. putrefaciens 200 was found to express low-rate nitrate reductase, nitrite reductase, and ferrireductase activity after growth under highly aerobic conditions and greatly elevated rates of each reductase activity after growth under microaerobic conditions. The effects of NO3- and NO2- on the Fe3+ reduction activity of both aerobically and microaerobically grown cells appeared to follow a consistent pattern; in the presence of Fe3+ and either NO3- or NO2-, dissimilatory Fe3+ and nitrogen oxide reduction occurred simultaneously. Nitrogen oxide reduction was not affected by the presence of Fe3+, suggesting that S. putrefaciens 200 expressed a set of at least three physiologically distinct terminal reductases that served as electron donors to NO3-, NO2-, and Fe3+. However, Fe3+ reduction was partially inhibited by the presence of either NO3- or NO2-. An in situ ferrozine assay was used to distinguish the biological and chemical components of the observed inhibitory effects. Rate data indicated that neither NO3- nor NO2- acted as a chemical oxidant of bacterially produced Fe2+. In addition, the decrease in Fe3+ reduction activity observed in the presence of both NO3- and NO2- was identical to the decrease observed in the presence of NO2- alone. These results suggest that bacterially produced NO2- is responsible for inhibiting electron transport to Fe3+.  相似文献   

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Summary An automated assay system was used to screen inhibitors of sulfide formation by sulfate-reducing bacteria. The system used a Technicon AutoAnalyzer II instrument both as an incubation chamber (to combine bacterial cells, substrates, and test compounds) and as an analyzer of the amount of sulfide formed during incubation. A lactate-limited chemostat was used to provide a reproducible inoculum of Desulfovibrio desulfuricans cells. The assay was useful in detecting even small changes in the rate of sulfate reduction resulting from exposure to inhibitors.Contribution no. 5652 from the Central Research and Development DepartmentOffprint requests to: P. J. Weimer  相似文献   

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A new strain of sulphate reducing bacteria was isolated from swampy forest soil. After 120 h reduction, sulphate conversion attained 100%, the molar ratio of the consumed lactate and reduced sulphate amounted to 2:1. This confirms the reduction mechanism proposed by Senez (1951).  相似文献   

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Studies on nitrate reduction by Escherichia coli   总被引:3,自引:1,他引:3       下载免费PDF全文
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Intensive agriculture leads to increased nitrogen fluxes (mostly as nitrate, NO3 ?) to aquatic ecosystems, which in turn creates ecological problems, including eutrophication and associated harmful algal blooms. These problems have focused scientific attention on understanding the controls on nitrate reduction processes such as denitrification and dissimilatory nitrate reduction to ammonium (DNRA). Our objective was to determine the effects of nutrient-tolerant bioturbating invertebrates (tubificid oligochaetes) on nitrogen cycling processes, specifically coupled nitrification–denitrification, net denitrification, DNRA, and biogeochemical fluxes (O2, NO3 ?, NH4 +, CO2, N2O, and CH4) in freshwater sediments. A mesocosm experiment determined how tubificid density and increasing NO3 ? concentrations (using N15 isotope tracing) interact to affect N cycling processes. At the lowest NO3 ? concentration and in the absence of bioturbation, the relative importance of denitrification to DNRA was similar (i.e., 49.6 and 50.4 ± 8.1 %, respectively). Increasing NO3 ? concentrations in the control cores (without fauna) stimulated denitrification, but did not enhance DNRA, which significantly altered the relative importance of denitrification compared to DNRA (94.6 vs. 5.4 ± 0.9 %, respectively). The presence of tubificid oligochaetes enhanced O2, NO3 ?, NH4 + fluxes, greenhouse gas production, and N cycling processes. The relative importance of denitrification to DNRA shifted towards favoring denitrification with both the increase in NO3 ? concentrations and the increase of bioturbation activity. Our study highlights that understanding the interactions between nutrient-tolerant bioturbating species and nitrate contamination is important for determining the nitrogen removal capacity of eutrophic freshwater ecosystems.  相似文献   

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1. A method is described for preparing spheroplasts from Paracoccus denitrificans that are substantially depleted of dissimilatory nitrate reductase (cytochrome cd) activity. Treatment of cells with lysozyme + EDTA together with a mild osmotic shock, followed by centrifugation, yielded a pellet of spheroplasts and a supernatant that contained d-type cytochrome. The spheroplasts were judged to have retained an intact plasma membrane on the basis that less than 1% of the activity of a cytoplasmic marker protein, malate dehydrogenase, was released from the spheroplasts. In addition to a low activity towards added nitrite, the suspension of spheroplasts accumulated the nitrite that was produced by respiratory chain-linked reduction of nitrate. It is concluded that nitrate reduction occurs at the periplasmic side of the plasma membrane irrespective of whether nitrite is generated by nitrate reduction or is added exogenously. 2. Further evidence for the integrity of the spheroplasts was that nitrate reduction was inhibited by O2, and that chlorate was reduced at a markedly lower rate than nitrate. These data are taken as evidence for an intact plasma membrane because it was shown that cells acquire the capability to reduce nitrate under aerobic conditions after addition of low amounts of Triton X-100 which, with the same titre, also overcame the permeability barrier to chlorate reduction by intact cells. The close relationship between the appearance of chlorate reduction and the loss of the inhibitory effect of O2 on nitrate reduction also suggests that the later feature of nitrate respiration is due to a control on the accessibility of nitrate to its reductase rather than on the flow of electrons to nitrate reductase.  相似文献   

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Wetlands are often highly effective nitrogen (N) sinks. In the Lake Waco Wetland (LWW), near Waco, Texas, USA, nitrate (NO3) concentrations are reduced by more than 90% in the first 500 m downstream of the inflow, creating a distinct gradient in NO3 concentration along the flow path of water. The relative importance of sediment denitrification (DNF), dissimilatory NO3 reduction to ammonium (DNRA), and N2 fixation were examined along the NO3 concentration gradient in the LWW. “Potential DNF” (hereafter potDNF) was observed in all months and ranged from 54 to 278 μmol N m−2 h−1. “Potential DNRA” (hereafter potDNRA) was observed only in summer months and ranged from 1.3 to 33 μmol N m−2 h−1. Net N2 flux ranged from 184 (net denitrification) to −270 (net N2 fixation) μmol N m−2 h−1. Nitrogen fixation was variable, ranging from 0 to 426 μmol N m−2 h−1, but high rates ranked among the highest reported for aquatic sediments. On average, summer potDNRA comprised only 5% (±2% SE) of total NO3 loss through dissimilatory pathways, but was as high as 36% at one site where potDNF was consistently low. Potential DNRA was higher in sediments with higher sediment oxygen demand (r 2 = 0.84), and was related to NO3 concentration in overlying water in one summer (r 2 = 0.81). Sediments were a NO3 sink and accounted for 50% of wetland NO3 removal (r 2 = 0.90). Sediments were an NH4+ source, but the wetland was often a net NH4+ sink. Although DNRA rates in freshwater wetlands may rival those observed in estuarine systems, the importance of DNRA in freshwater sediments appears to be minor relative to DNF. Furthermore, sediment N2 fixation can be extremely high when NO3 in overlying water is consistently low. The data suggest that newly fixed N can support sustained N transformation processes such as DNF and DNRA when surface water inorganic N supply rates are low.  相似文献   

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The strictly anaerobic Archaeon Ferroglobus placidus was grown chemolithoautotrophically on H2 and nitrate and analyzed for enzymes and coenzymes possibly involved in autotrophic CO2 fixation. The following enzymes were found [values in parentheses = μmol min–1 (mg protein)–1]: formylmethanofuran dehydrogenase (0.2), formylmethanofuran:tetrahydromethanopterin formyltransferase (0.6), methenyltetrahydromethanopterin cyclohydrolase (10), F420-dependent methylenetetrahydromethanopterin dehydrogenase (1.5), F420-dependent methylenetetrahydromethanopterin reductase (0.4), and carbon monoxide dehydrogenase (0.1). The cells contained coenzyme F420 (0.4 nmol/mg protein), tetrahydromethanopterin (0.9 nmol/ mg protein), and cytochrome b (4 nmol/mg membrane protein). From the enzyme and coenzyme composition of the cells, we deduced that autotrophic CO2 fixation in F. placidus proceeds via the carbon monoxide dehydrogenase pathway as in autotrophically growing Archaeoglobus and Methanoarchaea species. Evidence is also presented that cell extracts of F. placidus catalyze the reduction of two molecules of nitrite to 1 N2O with NO as intermediate (0.1 μmol N2O formed per min and mg protein), showing that – at least in principle –F. placidus has a denitrifying capacity. Received: 23 August 1996 / Accepted: 6 November 1996  相似文献   

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A membrane-bound nitrate reductase (nitrite:(acceptor) oxidoreductase, EC 1.7.99.4) from the extremely halophilic bacterium Haloferax denitrificans was solubilized by incubating membranes in buffer lacking NaCl and purified by DEAE, hydroxylapatite, and Sepharose 6B gel filtration chromatography. The purified nitrate reductase reduced chlorate and was inhibited by azide and cyanide. Preincubating the enzyme with cyanide increased the extent of inhibition which in turn was intensified when dithionite was present. Although cyanide was a noncompetitive inhibitor with respect to nitrate, nitrate protected against inhibition. The enzyme, as isolated, was composed of two subunits (Mr 116,000 and 60,000) and behaved as a dimer during gel filtration (Mr 380,000). Unlike other halobacterial enzymes, this nitrate reductase was most active, as well as stable, in the absence of salt.  相似文献   

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The processes involved in nitrate metabolism in Halobacterium of the Dead Sea are part of a dissimilatory pathway operating in these bacteria. The induction of both nitrate and nitrite reductases is shown to depend on the presence of nitrate and of anaerobic conditions. The gas products of the denitrification process were identified as nitrous oxide and nitrogen. Some properties of two of the enzymes involved in this process, nitrate and nitrite reductases, are reported. It is shown that the 2 Feferredoxin, which is present in large quantities in Halobacterium of the Dead Sea, can serve as an electron donor for nitrite reduction by nitrite reductase. It is suggested that the presence of a dissimilatory pathway for the reduction of nitrate in Halobacterium of the Dead Sea can be used as a tool for its classification.  相似文献   

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